SID-5 Is an Endosome-Associated Protein Required for Efficient Systemic RNAi in C. elegans

In the nematode C. elegans, RNAi silencing signals are efficiently taken up from the environment and transported between cells and tissues [1–3]. Previous studies implicating endosomal proteins in systemic RNAi lack conclusive evidence [4, 5]. Here, we report the identification and characterization of SID-5, a C. elegans endosome-associated protein that is required for efficient systemic RNAi in response to both ingested and expressed double-stranded RNA (dsRNA). SID-5 is detected in cytoplasmic foci that partially colocalize with GFP fusions of late endosomal proteins RAB-7 and LMP-1. Furthermore, knockdown of various endosomal proteins similarly relocalizes both SID-5 and LMP-1::GFP. Consistent with a non-cell-autonomous function, intestine-specific SID-5 expression restored body wall muscle (bwm) target gene silencing in response to ingested dsRNA. Finally, we show that sid-5 is required for the previously described sid-1-independent transport of ingested RNAi triggers across the intestine [6]. Together, these data demonstrate that an endosome-associated protein, SID-5, promotes the transport of RNAi silencing signals between cells. Furthermore, SID-5 acts differently than the previously described SID-1, SID-2, and SID-3 proteins [3, 6–8], thus expanding the systemic RNAi pathway. ► sid-5 mutants are defective in systemic RNAi ► The SID-5 protein colocalizes with late endosomal proteins RAB-7 and LMP-1 ► Intestinal SID-5 overexpression restores silencing in muscle cells in a sid-5 mutant ► sid-5 is required for sid-1-independent intestinal transport of silencing signals